去泛素化酶PSMD7对膀胱癌增殖和迁移的影响  

Impact of deubiquitinating enzyme PSMD7 on bladder cancer proliferation and migration

作　　者：余力 谢冲 丁勇杰 刘涛[3] 朱卫[2] 李艳松 胡祎舜 刘敏豪 汤晨昊 刘子阳 彭袈豪 袁作洈 柳栩然 曾金敏[1] 廖义翔[1] Yu Li;Xie Chong;Ding Yongjie;Liu Tao;Zhu Wei;Li Yansong;Hu Yishun;Liu Minhao;Tang Chenhao;Liu Ziyang;Peng Jiahao;Yuan Zuowei;Liu Xuran;Zeng Jinmin;Liao Yixiang(Department of Urology,Jingzhou Central Hospital,Jingzhou Hospital Affiliated to Yangtze University,Jingzhou 434020,China;Department of Urology,Jiangling County People’s Hospital,Jiangling 434100,China;Department of Urology,Wuhan University Zhongnan Hospital,Wuhan 430071,China)

机构地区：[1]荆州市中心医院长江大学附属荆州医院泌尿外科,荆州434020 [2]江陵县人民医院泌尿外科,江陵434100 [3]武汉大学中南医院泌尿外科,武汉430071

出　　处：《中华实验外科杂志》2024年第5期1007-1010,共4页Chinese Journal of Experimental Surgery

摘　　要：目的:探讨去泛素化酶PSMD7的表达对膀胱癌增殖与迁移的影响。方法:通过Timer数据库分析PSMD7在膀胱癌中的差异表达。在膀胱癌细胞株(UMUC3和T24)中使用成簇的规律间隔短回文重复序列(CRISPR)/Cas9技术导入短发夹RNA(shRNA)敲低PSMD7。设置组别为对照组和稳定敲低PSMD7实验组UMUC3(UM-sh-1组和UM-sh-2组)和T24(T24-sh-1组和T24-sh-2组)。通过细胞计数试剂盒(CCK-8)实验、平板克隆形成实验、Transwell实验观察PSMD7对膀胱癌增殖与迁移的影响。通过免疫荧光实验检测细胞增殖标志物5-乙炔基-2’-脱氧尿苷(EDU)验证敲低PSMD7抑制膀胱癌细胞的增殖。应用独立样本t检验进行组间比较。结果:Timer数据库发现PSMD7在多种癌症中高表达(P<0.05),其中包括膀胱癌,肿瘤组织中PSMD7的表达高于正常组织,差异有统计学意义[8.3(7.5~9.1)比3.1(2.2~4.1),P<0.05]。CCK-8结果显示,敲低PSMD7后细胞增殖能力明显低于对照组[UMUC3细胞株中对照组吸光度值明显高于敲低组(1.724±0.057比0.879±0.040,t=20.83,P<0.05)和(1.724±0.057比0.935±0.065,t=15.62,P<0.05);T24细胞株中对照组吸光度值明显高于敲低组(2.329±0.084比0.973±0.072,t=21.14,P<0.05)和(2.329±0.084比0.830±0.087,t=21.41,P<0.05)],平板克隆形成实验结果显示,PSMD7敲低后细胞增殖能力明显低于对照组[UMUC3细胞株中对照组集落数明显高于敲低组(716.5±18.5)个比(508.0±21.0)个,t=7.45,P<0.05和(716.5±18.5)个比(396.5±9.5)个,t=15.39,P<0.05;T24细胞株中对照组集落数明显高于敲低组(687.0±33.0)个比(359.5±15.5)个,t=8.983,P<0.05和(687.0±33.0)个比(407.0±18.0)个,t=7.449,P<0.05]。Transwell实验显示,PSMD7敲低后细胞迁移能力显著低于对照组[UMUC3细胞株中对照组迁移数明显高于敲低组(756.0±28.0比198.0±9.5,t=18.86,P<0.05)和(756.0±28.0比146.5±11.5,t=20.14,P<0.05);T24细胞株中对照组迁移数明显高于敲低组[(648.0±18.0)个比(132.0±7.0)个,t=25.79,P<0.05和(648.0±Objective To investigate the effect of deubiquitinating enzyme PSMD7 expression on bladder cancer proliferation and migration.Methods The differential expression of PSMD7 in bladder cancer was analyzed by Timer database.In bladder cancer cell lines(UMUC3 and T24),regular interval short palindromic repeat clusters(CRISPR)/Cas9 technology was used to introduce short hairpin RNA(shRNA)to knock down PSMD7.The following groups were set up:control group and stable PSMD7 knockout experimental groups[UMUC3(UM-sh-1 group and UM-sh-2 group)and T24(T24-SH-1 group and T24-SH-2 group)].The effects of PSMD7 on the proliferation and migration of bladder cancer cells was investigated by cell counting kit-8(CCK-8)assay,plate clone formation assay and Transwell assay.Cell proliferation marker 5-acetylidene-2’-deoxyuridine(EDU)was detected by immunofluorescence assay to verify that PSMD7 knockdown inhibited the proliferation of bladder cancer cells.The independent sample t test was used for comparison between groups.Results The Timer database found that PSMD7 was highly expressed in a variety of cancers(P<0.05),including bladder cancer,and the expression of PSMD7 in tumour tissues was higher than that in normal tissues,with a statistically significant difference[8.3(7.5-9.1)vs.3.1(2.2-4.1),P<0.05].The CCK-8 results showed that the proliferative capacity of the cells was significantly decreased after knockdown of PSMD7 as compared with the control group[the absorbance of the control group was significantly higher than that of the knockdown group in the UMUC3 cell line(1.724±0.057 vs.0.879±0.040,t=20.83,P<0.05)and(1.724±0.057 vs.0.935±0.065,t=15.62,P<0.05);The absorbance of the control group was significantly higher than that of the knockdown group in the T24 cell line(2.329±0.084 vs.0.973±0.072,t=21.14,P<0.05)and(2.329±0.084 vs.0.830±0.087,t=21.41,P<0.05)].The results of the plate clone formation assay showed that cell proliferation ability was significantly decreased after PSMD7 knockdown[The number of colonies in the con

关 键 词：膀胱癌 去泛素化酶 增殖 迁移 

分 类 号：R737.14[医药卫生—肿瘤]